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Development of a Real-Time Quantitative PCR Assay to Detect Cryptosporidium parvum Oocysts in Soil
Citation:
Liang, Z. AND A. AZADPOUR-KEELEY. Development of a Real-Time Quantitative PCR Assay to Detect Cryptosporidium parvum Oocysts in Soil. Presented at 9th Annual Biodefense and Emerging Diseases ResearchMeeting, Washington, DC, February 05 - 09, 2011.
Impact/Purpose:
Poster for the 9th Biodefense and Emerging Diseases Research Meeting, American Society for Microbiology, Washington, DC (February 5-9, 2011)
Description:
The risk of Cryptosporidium parvum (C. parvum) contamination is a serious issue with respect to drinking water, as evidenced by the cryptosporidiosis outbreak in Milwaukee WI, in 1993, which involved over 400,000 infections and at least 54 deaths. Ground-water contamination by C. parvum either directly or through infiltration may also present a challenge since ground water serves as the source of drinking water for roughly half of the domestic needs of the United States. Contaminated soil can serve as an important source and pathway from which Cryptosporidium oocysts can be transported into water sources. Therefore, rapid and sensitive techniques are required for the efficient detection of C. parvum in soil. The currently used Immunomagnetic separation (IMS) method (USEPA method 1623) is slow, expensive and inefficient to separate C. parvum from environmental sample. Real time PCR offers a more rapid, sensitive and precise alternative. Most of the available PCR-based methods, however, were developed for clinical samples and because of the differences in density of oocysts in environmental samples as well as matrix effects their applications are questionable. There is a need to develop detection procedures that specifically target soil samples. It has been shown that DNA extraction techniques may affect the sensitivity of Real time PCR procedures. Therefore, the selection of a proper extraction method for a given soil samples is paramount. This poster does not necessarily reflect EPA policy. Mention of trade names or commercial products does not constitute endorsement or recommendation for use.
